Abstract 1249: MLN4924, a protein neddylation inhibitor, induces apoptosis via endoplasmic reticulum (ER)-stress in human cervical cancer

Abstract

Abstract Cervical cancer is the second most common cancer among the women worldwide. The treatment of the cervical cancer includes the surgery, chemotherapy, external and intracavitary radiotherapy adjusted to the different cancer stage. Despite these treatments, a certain proportion of women still die of the cancer due to recurrent or chemoresistent disease. The toxicity and side effect of these therapies are substantial. Therefore, it is important to develop new drugs or combination treatments to improve patient survival and minimize side effect of the therapy. MLN4924 is an inhibitor of NEDD8 activating enzyme (NAE), has been reported to elicit anti-tumor response in various malignancies. In this study, we aim to investigate the anti-tumor effect of MLN4924 on human cervical cancer and underlying mechanism in vitro and in vivo. We used two different grade human bladder cervical cell lines: ME-180 and HeLa. After exposing to MLN4924, the cell viability and apoptosis were measured by MTT and flow cytometry with annexin V-FITC labeling. The efficacy of cell proliferation was determined by BrdU incorporation assay. The cell cycle regulators, cell death-related molecules, and ER stress-related proteins were examined by Western blotting. We also use Nu/Nu xenograft mice model to clarify the effect of MLN4924 on tumor growth in vivo. We demonstrated that MLN4924 effectively induces dose-dependent cytotoxicity, apoptosis and anti-tumor phenomenon in cervical cancer cells. MLN4924 also activate cleaved-Bid, phospho-histone H2A.X, caspases and PARP, decresed phospho-Bcl2, and cause cell cycle arrest. Furthermore, MLN4924 possessed the ability to activate ER stress-related moleculors caspase-4, and CCAAT/enhancer binding protein homologous protein (CHOP), which were involved in UC cell apoptosis, and other stress responses (JNK and c-Jun activations). Moreover, we confirmed MLN4924 inhibited tumor growth and induced ER-stress in a cervical cancer xenograft mice model. Taken together, we demonstrated that MLN4924 induces cytotoxicity and cell cycle retardation, as well as activation of ER stress responses in human cervical cancer. The anti-tumor effect of MLN4924 provides a novel implication in clinical treatment of cervical cancer. Citation Format: Kuan-Lin Kuo, Yeong-Shiau Pu, I-LIN Ho, Chen-Hsun Hsu, Ju-Ton Hsieh, Wei-Chou Lin, Chien-Tso Chou, Kuo-How Huang. MLN4924, a protein neddylation inhibitor, induces apoptosis via endoplasmic reticulum (ER)-stress in human cervical cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1249. doi:10.1158/1538-7445.AM2015-1249